树鼩慢性感染乙肝病毒过程中枯否细胞变化的意义

Changes of Kupffer cells during tree shrew chronically infected with hepatitis B virus

目的:探索枯否细胞在树鼩感染乙肝病毒(HBV)慢性化过程中的意义。方法:树鼩分为3组:A组6只,为前期实验已确定慢性感染HBV的树鼩;B组3只,为疑似慢性感染HBV的树鼩;C组4只,为未接种HBV的正常对照树鼩。全部动物定期抽血和进行肝活检手术;对手术切取的树鼩肝组织进行枯否细胞的分离、纯化和原代培养,采用流式细胞术、细胞免疫组化、溶酶体荧光探针及实时荧光定量RT-PCR等方法检测CD163+细胞数量、溶酶体数量、溶菌酶的表达及肿瘤坏死因子α(TNF-α)mRNA表达水平。结果:(1)慢性感染HBV的树鼩肝脏枯否细胞比例及肝组织内CD163+细胞数量显著高于其它2组(均P<0.05);(2)慢性感染HBV的树鼩肝脏枯否细胞的溶酶体荧光强度、肝组织内溶菌酶阳性细胞计数和TNF-αmRNA的表达水平均显著低于其它2组(均P<0.05)。结论:枯否细胞在宿主感染HBV的慢性化过程中可能起一定的调节作用。

AIM：To explore the changes and significance of Kupffer cells in the process of tree shrew chroni -cally infected with hepatitis B virus （HBV）.METHODS：The animals were divided into 3 groups.Group A consists of 6 tree shrews that were identified as persistently infected with HBV;group B consists of 3 tree shrews that were suspected as persistently infected with HBV;group C consists of 4 tree shrews that were not inoculated with HBV and were applied as normal controls.Liver biopsies were collected regularly from all animals , and the Kupffer cells were isolated , purified and primarily cultured.The techniques of flow cytometry , immunohistochemistry, lysosomal fluorescent probe staining and real-time RT-PCR were applied to determine the number and function of these Kupffer cells .RESULTS： The result showed that the count and proportion of CD 163^＋cells in group A were significantly higher than those in group B and group C （ P〈0.05）.Meanwhile, the fluorescence intensity levels of lysosomal , the number of lysozyme-positive cells and the mRNA expression level of TNF-αin the Kupffer cells in group A were significantly lower than those in group B and group C （ P〈0.05）.CONCLUSION：Kupffer cells may play a regulatory role during host’s chronic HBV infection.